Kun:LabNotes/ASE/2007-11-10
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Exp.1 Probe prep. for the CES22k set
In this round of probe prep, I'm making probes from three different batches of Agilent oligos.
- CES22k (from Harvard, has been sequenced)
- CES22k 01783910026 (received Oct. 2007, same sequences as the above library)
- CES22k 10783810002 (received Oct. 2007, contains Solexa amplification adaptors)
Emily told me tube #2 and #3 should have 10pmol of DNA, so I'm resuspending the oligos in 100ul nuclease free H2O for the concentration of 100nM.
PCR (with iTaq)
x1 x16x3 template(100nM) 0.1ul 3x1.6ul 10x ABI buffer 10ul 480ul 25mM MgCl2 6ul 288ul 10mM dNTP 2ul 96ul 100uM pAP1V6U 0.3ul 14.4ul 100uM phosAP2V6 0.3ul 14.4ul 50X SYBG I 0.8ul 38.4ul iTaq 1ul 48ul H2O 80ul 3840ul
94C 3min -> 18 cycles of (94C 45sec -> 58C 2min -> 72C 1min) -> 72C 3min -> 15C hold